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Starting Material: DNA

Submitting Samples for DNA-Seq Library Prep & Sequencing

  1.  Consult with us on your project before making any requests. 
  2.  Determine which bulk DNA library preparation fits your needs.
    1. See the tabs below for sample concentration and volume requirements. 
  3.  Download our form, fill in all highlighted fields, and submit your form through minilims.
  4.  Array samples by column in a semi or full-skirted 96-well plate.
  5. Place samples in a provided zip-top bag with the completed submission slip.
  6.  Log your sample drop-off on the paper log.
  7.  Place the bag into the “sequencing drop-off" submission freezer in the hall.

Note: See here if you only want sequencing services.

ngs_dna_library_service_request_form_fy24.xlsx598 KB

DNA Library Preparations

1/4 Kapa DNA HyperPlus- Ligation Based

Using a Mantis Low Volume Liquid Dispenser (Formulatrix) and a Biomek FXP Span-8 Liquid Handler (Beckman Coulter) the Core will prepare sequencing libraires using the Kapa HyperPlus Kit (Roche Sequencing). 
 
  • Enzymatic DNA fragmentation and Adapter Ligation
  • Compatible with amplicons, already fragmented gDNA, highly-degraded gDNA, and low-input samples
  • Optional library amplification using low-bias PCR 
  • Unique dual indexes
 
To submit samples for Kapa HyperPlus Library Preparation, please provide the following:
  • 15 ul of DNA between 1-10 ng/ul
    • Core will perform fluorometric assessment of input sample to verify concentration
    • Core can normalize input sample as a service
    • Samples below 8.75 ng input are not guaranteed (we have seen success with lower inputs)
  • Note: If your samples are intact and not highly-degraded DNA, we recommend either the Illumina DNA Libray Prep Kit.
Image of 96-well plate with arrow down the first columnOther requirements:
  • Samples must be arrayed in columns of a 96-well plate 
  • Samples and all appropriate Minilims requests must be submitted by the 15th of the month (or the Friday before as applicable)
    • If the core is completing QC on input samples submission date is the 8th of the month
    • Turnaround time is 4 weeks from the 15th of the month
    • Sequencing is ~2 weeks, but is dependent on platform and run parameters
  • Drop-off samples in the freezer outside the core between 7 AM- 7 PM
 

1/4 Nextera XT- Tagmentation

Using a Mantis Low Volume Liquid Dispenser (Formulatrix) and a Biomek FXP Span-8 Liquid Handler (Beckman Coulter) the Core will prepare sequencing libraires using the Nextera XT DNA Library Prep Kit (Illumina). 
 
  • Transposase-mediated tagmentation based library prep
    • Tagmentation: gDNA is fragmented and adapter sequences added
  • Combinatorial dual indexes are incorporated via PCR amplification  
 
To submit samples for Illumina DNA Library Preparation, please provide the following:
  • 10 ul of DNA between 0.15-0.2 ng/ul (maximum input: 0.25 ng)
    • Core will perform fluorometric assessment of input sample to verify concentration
    • Core can normalize input sample as a service
    • Samples below ~0.15 ng input are not guaranteed 
  • Note: If your samples are highly-degraded DNA, already fragmented before submission, or are amplicons, we recommend the Kapa DNA HyperPlus Kit
Image of 96-well plate with arrow down the first columnOther requirements:
  • Samples must be arrayed in columns of a 96-well plate 
  • Samples and all appropriate Minilims requests must be submitted by the 1st of the month (or the Friday before as applicable)
    • If the core is completing QC on input samples submission date is the 25th of the previous month
    • Turnaround time is 4 weeks from the 1st of the month
    • Sequencing is ~2 weeks, but is dependent on platform and run parameters
  • Drop-off samples in the freezer outside the core between 7 AM- 7 PM

1/4 Illumina DNA- On Bead Tagmentation

Using a Mantis Low Volume Liquid Dispenser (Formulatrix) and a Biomek FXP Span-8 Liquid Handler (Beckman Coulter) the Core will prepare sequencing libraires using the Illumina DNA Prep Kit (Illumina). 
 
  • Transposase-mediated tagmentation based library prep
    • Using on bead tagmentation gDNA is fragmented and adapter sequences added
  • Unique dual indexes are incorporated via PCR amplification  
 
To submit samples for Illumina DNA Library Preparation, please provide the following:
  • 15 ul of DNA between 0.07-16.5 ng/ul
    • Core will perform fluorometric assessment of input sample to verify concentration
    • Core can normalize input sample as a service
    • Samples below 0.5 ng input are not guaranteed 
  • Note: If your samples are highly-degraded DNA, already fragmented before submission, or are amplicons, we recommend the Kapa DNA HyperPlus Kit
Image of 96-well plate with arrow down the first columnOther requirements:
  • Samples must be arrayed in columns of a 96-well plate 
  • Samples and all appropriate Minilims requests must be submitted by the 1st of the month (or the Friday before as applicable)
    • If the core is completing QC on input samples submission date is the 25th of the previous month
    • Turnaround time is 4 weeks from the 1st of the month
    • Sequencing is ~2 weeks, but is dependent on platform and run parameters
  • Drop-off samples in the freezer outside the core between 7 AM- 7 PM